Background and objectives: The immunobead binding TEST (IBT) and the mixed AGGLUTINATION reaction (MAR) are the most commonly used methods for detection of antisperm antibodies (ASA). The detection of ASA by flow cytometry (FCM) was first described by Haas and Cunningham. Both assays can be performed as DIRECT or inDIRECT methods. In this study, inDIRECT FCM was compared with the DIRECT MAR for detection of ASA. Methods: Semen samples were obtained from 80 men (infertile couples) in Isfahan Fertility and Infertility Center. Seminal plasma samples were incubated with ASA-negative donor sperm. Then, surface-bound antibody was detected with FITC-labeled antihuman immunoglobulin DIRECTed against IgA and IgG in the inDIRECT FCM assay. ASAs bound to the surface of patients’ sperm were detected by DIRECT MAR TEST. Results: The inDIRECT FCM correlates with DIRECT MAR for detection of IgA antisperm antibodies (r=0.55 and P=0.006). The inDIRECT FCM, however, does not correlate with DIRECT MAR for the detection of IgG antisperm antibodies (r=0.25 and P=0.25). Conclusion: Some of the ASAs in seminal fluid bind to spermatozoa. Therefore, inDIRECT TESTs to detect ASAs in seminal plasma are likely to miss the presence of IgG antisperm antibodies while they effectively detect IgA antisperm antibodies.

Background: This study aimed to set-up latex AGGLUTINATION TEST (LAT) and ELISA based on recombinant A2 from Iranian strain of Leishmania (L. ) infantum (rA2-Ag) and evaluated for detection of anti-Leishmania antibodies in dogs compared to standard DIRECT AGGLUTINATION TEST (DAT). Methods: The rA2-Ag was synthesized under a part of the A2 gene sequences which contain immune dominant sequences and less number of repetitive sequences. Latex beads, 0. 8 μ m (Sigma, USA) were sensitized with rA2-Ag. The TESTs were carried out on sera collected from 350 ownership dogs including symptomatic (n=67), asymptomatic (n=230) canine visceral leishmaniasis (CVL), and (n=53) uninfected domestic dogs as control group. Results: Anti-leishmanial antibodies were detected in 97 (27. 7%), 96 (27. 4%) and 29 (%9) of the serum samples by using DAT, rA2-ELISA, and rA2-latex, respectively with ≥ 1: 320 as a cut-off titer when DAT-confirmed cases were compared with the control groups. A combined sensitivity of 52% and specificity of 82. 40% for rA2-ELISA and 23. 8% and specificity 95. 38%, respectively were found with ≥ 1: 320 as a cut-off titer when DAT-confirmed cases were compared with the control groups. The concordance between rA2-ELISA and rA2 latex compared with DAT as a gold standard serological TEST for VL were found 73. 7% and 77. 5%, respectively. Conclusion: A good degree of agreement was found between rA2-ELISA and DAT (73. 7%). rA2-ELISA could detect more seropositive serum samples than rA2-LAT and it may be recommended as an alternative tool for the diagnosis of CVL.

Background: Visceral leishmaniasis (VL) or Kala azar is an infectious disease caused by various species of Leishmania parasites. The aim of this study was to detect and compare the presence of anti-Leishmania antibodies in the urine of visceral leishmaniasis patients using ELISA and DAT methods.Methods: A total of 30 urine samples were collected from VL patients referred to Shiraz (southeast of Iran) hospitals. Moreover 31 urine samples were collected from healthy individuals and patients with other diseases such as malaria, brucellosis, hydatidosis and cutaneous leishmaniasis. Collected samples were examined to detect anti-Leishmania antibodies in urine, using ELISA and DAT.Results: Anti-Leishmania antibody was detected in urine of 18 out of 30 (60%) VL patients by DAT while ELISA detected anti-Leishmania antibodies in urine of 28 out of 30 (93.3%) of VL cases. Sensitivity and specificity of urine-based DAT was 60% and 83.9%, respectively while sensitivity and specificity of urine-based ELISA were 93.3% and 93.5%, correspondingly.Conclusion: Urine-based DAT and ELISA have a reasonable specificity and sensitivity in diagnosis of VL. Accordingly, urine-based ELISA might be a suitable alternative for serum based assays for diagnosis of VL.

Background & Objectives: This study aimed to determine of the seroprevalence of visceral leishmaniasis to design a prevention and control program in Bojnoord district.Methods: This was a descriptive and cross sectional study with multistage random sampling method. The study was carried out on children up to 12 years old and 10% of adult’s population in 8 rural areas of Bojnoord district in 2007. In total, 1608 blood samples were collected to detect anti-Leishmania antibodies. All the samples were TESTed by DIRECT AGGLUTINATION TEST (DAT). In order to determine Leishmania species, necropsy was performed on four suspected dogs in the studied areas. The species of Leishmania was determined by RAPD-PCR and PCR-RFLP methods using to amplify the ribosomal internal transcribed spacer 1 (ITS1).Results: Thirty and eight (2.36%) out of 1608 collected blood samples had anti-Leishmania antibodies at titer 1:800 and nine (0.56%) were just positive at 1:3200 by DAT. There was no statistically significant difference between female and male seroprevalence (p>0.05). The seoprevalence in children <=12 years old compared to individuals greater than 12 years old did not show any statistically significant (p>0.05). Amastigotes were observed in all 4 necropsied dogs. The species of Leishmania isolated from 2 dogs, was determined as L.Infantum. Their ITS1 sequences were registered with Accession numbers EU810776 and EU810777 in NCBI.Conclusions: These findings showed that visceral leishmaniasis has been circulated with low endemicity in Bojnoord district. Therefore an appropriate monitoring system is needed for health services in this area.

Visceral leishmaniasis (VL) is one of the most important parasitic diseases which are endemic in different parts of Iran. Serological studies were conducted by DIRECT AGGLUTINATION TEST (DAT) on 12144 human serum samples, collected from four geographical zones of Iran. Sero prevalence, geographical distribution, clinical signs and symptoms for human visceral leishmaniasis based on DAT for the period of 2002 through 2005 were determined. From 516 kala-azar cases detected: 50.6% were from Meshkin-shahr and Moghan districts in Ardabil Province, northwest of Iran and 49.4% were detected from other areas of Iran. In physical examination of seropositive cases, which were detected by DAT with anti leishmanial antibodies at titers of 1: 3200 to 1: 102400, almost 50% of suspected individuals showed the classical kala-azar signs and symptoms. Predominant signs and symptoms in 233 hospitalized patients with anti-Leishmania antibodies at 1:3200 and higher, were fever (88.0%) and splenomegaly (84.5%). Statistically significant difference was found between males (58%) and females (42%) (P< 0.01). Moreover, 93.6% of the VL patients were £ 5 yr of age, and 6.4% were older than 5 yr that this difference was statistically significant (P< 0.01). From 1383 serum samples collected from domestic dogs in the villages that are known as endemic foci of human leishmaniasis, 152 (11.0%) were positive by DAT (³ 1:320). Parasitological and serological examinations that were performed in 30 wild canines showed that 10% of these animals were infected by L. infantum. L. infantum Lon 49 is the principal agent of the disease in human as well as animal reservoir hosts in different parts of Iran. For the first time in Iran, L. tropica isolated from both skin lesions in the face and bone marrow aspiration in a HIV+ man who co infected with VL as well as in an infected dog from Ardabil Province.

Background and purpose: Human visceral leishmaniasis (HVL) is endemic in several foci in IRAN, such as Ardebil and Fars provinces (in North western and southern parts of IRAN) and in some regions as sporadic. Visceral leishmaniasis in Iran is Mediterranean type and the causative agent is Leishmania infantum and its main reservoir is dog. Material and methods: In this study DIRECT AGGLUTINATION TEST (DAT) was compared with inDIRECT fluorescent antibody TEST (IFAT) for the diagnosis of visceral leishmaniasis in patients suspected of kala-azar. A total of 70 serum samples were collected from suspected kala-azar patients mainly in the kala-azar endemic areas. The Leishmania infantum antigens (MHO/TN/80/IPTi) were prepared in Department of parasitology, School of Medicine, Isfahan University of Medical Sciences. The principal phases of the procedure from making DAT antigen were mass production of promastigotes of leishmania in the RPMI1640 + fetal bovine serum, Trypsiniznation of parasites, staining with Comassie Blue and fixing with formaldehyde. The human serum samples were TESTed by DAT, as well as, by IFAT, with the L.infantum antigen prepared in our laboratory. Results: The sero positive rate (SPR) with DAT in titers of ≥ 1:3200 was 91.4% and with IFAT in titers of ≥ 1:80 was 94.3%. Discussion: Geometric means of reciprocal titers (GMRT) were 6309 for DAT and 692 for IFAT. Therefore, as the titers of ≥1:3200 are usually considered positive in DAT, the titers of ≥1:80 were regarded as position in IFAT. The coincidence of the two TESTs was 92%. These results showed that a simple local laboratory with one or two trained technicians is quite sufficient for DAT, sero-diagnosis and serological surveys of kala-azar in an endemic area. According to the results of this study, it seems that in Kala azar endemic areas, the clinical symptoms of Visceral leishmaniasis, particularly among the children with DAT antibody titers ≥1:3200 is a good indication for specific treatment of Kala-azar.    

Background & Objective: Visceral leishmaniasis (kala-azar) is an infectious-parasitic disease occurring sporadically in various parts of Iran. The disease is endemic in Ardabil, East Azarbaijan, Bushehr and Fars provinces. This study was performed with the aim of evaluation and comparison of DIRECT AGGLUTINATION TEST (OAT), inDIRECT immunofluorescent antibody (IFAT) and ELISA in sero-diagnosis of visceral leishmaniasis that is following a seroepidemiological survey in Germi district of Ardabil province.Methods: Cluster sampling has been performed from£12 years old and 10% of the adult population in Germi district. Altogether 1155 blood specimens were collected to detect anti- Leishmania antibodies. The samples were TESTed by DIRECT AGGLUTINATION TEST (OAT), inDIRECT immunofluorescent antibody (IFAT) and ELISA methods.Results: 32 (2.8%) of the collected specimens have anti-Leishmania antibodies with titers ³1:800 and from total specimens 7 (0.6%) were positive with ³1:3200 titers. In IFA TEST 32 (2.8%) have titers ³1:40 and from total specimens 6 (0.52%) were positive with ³1:320 titers. In ELISA TEST 8 specimens were positive and other specimens were negative.Conclusions: According to the results of this study, IFAT and ELISA are specific and high sensitive TESTs for sero-diagnosis of visceral leishmaniasis but these methods are expensive and require the complicated tools. Whereas, OAT is a simple method, not expensive and require a simple local laboratory with one or two trained technicians. Therefore OAT could be used for seroepidemiological studies and sero-diagnosis of visceral leishmaniasis in endemic areas instead of IFA and ELISA methods.

Comparison of serological results on 100 selected sera of sheep showed that qualitative correction between DIRECT AGGLUTINATION TEST (D.A) and dye TEST (D.T), DIRECT AGGLUTINATION TEST (D.A) and inDIRECT immunofluorescent antibody TEST (I.F.A.T), and dye TEST (D.T) and inDIRECT imunofluorescent TEST (LF.A.T) were 97%,96% and 99% respectively. Thus, in present study, it is concluded that maximal quantitative correlation was between DIRECT AGGLUTINATION TEST (D.A) and inDIRECT immunofluorescent antibody TEST (LF.A.T) and minimal quantitative correlation was between dye TEST (D.T) and inDIRECT immunofluorescent (I.F.A.T). Titration results of 100 sheep sera showed that the most frequent titers in DIRECT AGGLUTINATION (D.A), dye TEST (D.T) and inDIRECT Immunofluorescent (I.F.A.T) were 1/10240, 1/640, 1/1280 respectively. La this study, the geometric mean titers in DIRECT AGGLUTINATION TEST (D.A) was higher that two other serological methods.

Background: Leishmaniasis is an infection caused by a protozoan of the genus Leishmania. This descriptive and cross- sectional study was done in order to determine the seroepidemiology of visceral leishmaniasis in East Myankooh area, in Lorestan Province during 2004– 2005 time period.Materials and methods: This study was a descriptive, cross- sectional study. Multi stage sampling was performed from all children (<12 years old) and 10% 0f adults of this geographic region. Altogether, 530 blood samples were collected to detect antileishmania antibodies. The samples were TESTed by serological procedures including DIRECT AGGLUTINATION TEST (DAT). Antibody titers of 1:3200 and more were regarded as positive and 1:1600 titer was regarded as suspected. Findings: Six cases (1.26%) were serologically positive and 1 case (0.21%) was suspicious. The titer of antiserum in 6 cases (1.13%) was 1:800. Conclusion: Seropositive cases were seen in Latvan (n=1), Darsafeh (n=2) and Bungo (n=2) villages and in tribes (n=1). There were no cases of visceral leishmaniasis in other villages. This study indicated that visceral leishmaniasis is sporadic in East Myankooh. More complete epidemiological studies are necessary for determining vectors and other probable reservoirs in this region.